In order to properly wash slides; one needs a squeeze bottle, a Coplin jar and a slide holder. While slides are close to the end of their incubation period, the slide holder should be placed in the Coplin jar and then filled with fresh PBS. The squeeze bottle should then also be filled with PBS. The slides should be removed from the moist chamber, one at a time, and then the slides should be gently rinsed with the squeeze bottle PBS. The optimal way to perform this would be to direct the stream of PBS on the slide mask and not on the wells themselves. The slides should be tilted forward, gently rinsed and then tilted backwards and gently rinsed again. After this, the slides should have any excess liquid expelled from them by "snapping" the slide with a quick snap motion. Performing the gentle wash step in this format will reduce the possibility of well to well cross contamination. It is important to avoid aiming the PBS stream directly on the wells as this can dislodge substrate and interfere with results. Once this has been completed, the slide should be placed in the slide holder, in the Coplin jar and then the next consecutive slide should be processed in the same manner. Package insert specifications should be followed for each test system's respective wash requirement. Most IFA assays require a change of PBS in between wash incubations. This should be performed by removing the slide holder from the Coplin jar and then the old PBS should be removed and fresh PBS should be added to the Coplin jar. The slide holder should then be immediately replaced in the Coplin jar for their second wash incubation. Unless otherwise specified to dry slides after the wash procedure, the slides should remain in the Coplin jar filled with PBS and processed for conjugation or cover slipping individually, while remaining slides are still in PBS as to avoid drying. Slides that do not require drying after washing will contain a blotter in the slide packaging. After the required wash steps have been performed, the blotter should be laid on the lab bench (Note: avoid placing them on another absorptive surface) and then the slide should be lined up with the blotter so that the wells match up with the holes in the blotter. The actual well surface should never come in contact with the blotter or any other absorptive surface. Once the slide has been aligned, a paper towel should be used to dry the full glass, back portion of the slide. After this has been completed, the slide mask should be inspected for any additional moisture. If the slide mask appears dry, then the slides are ready to be conjugated or cover slipped depending on the next consecutive step in the Package Insert.

Negative results rely on the 1 to 4+ fluorescence grading system. 1+ will be the weakest fluorescence intensity that is considered reactive. While a sample may exhibit some fluorescence, positivity is dependent on its comparison to the 1+ fluorescence grade. Therefore, if the fluorescence is below the 1+ intensity the result would be considered negative.